16-keto and 16-hydroxy derivatives of 2-androsten-17beta-ol



United States Patent 3,243,445 IG-KETO AND 16-HYDROXY DERIVATIVES OF 2'-ANDR0STEN-17 8-'OL Max N. Huffman, Colorado Springs, (1010., assignor to v Lasdon Foundation, Inc., Yonkers, N.Y., a corporation 3 of Delaware No Drawing. Filed Apr. 7, 1964, Ser. No. 358,078

5 Claims. (Cl. 260-3973) This application is a continuation-in-part of my copending application Serial No. 249,616, filed January 7, 1963, now abandoned.

This invention relates to novel steroid compounds and to the production thereof. More particularly, this invention relates to novel steroids of the 2-androsten-17fl-ol series which have hydroxyl or keto substituents in the 16- position. These compounds have the following general structural formula wherein X represents a carbonyl group or a hydroxymethylene group.

The compounds of this invention are useful in the prevention and suppression of schizophrenic behavior induced in mammals by lysergic acid diet-hyl'amide (LSD). The peculiar psychic action of LSD was discovered by A. Hofmann and W. A. Stoll (Schweizer Archiv fur Neurologie and Psychiatrie, volume 60, page 279, 1947). Stoll showed that LSD produced an intoxication of the acute exogenous reaction type (as classified in psychic disease) in human beings when administered by mouth at the astonishingly small dose of 20 to micrograms.

G. Condrau demonstrated that schizophrenic subjects are much more resistant to the action of LSD than normal persons, and that the tolerance to LSD seemed as a rule better in psychopaths than in healthy individuals (Acta Psychiatrica at Neurologica Scandanavica, volume 24, page 9, 1949). Condrau raised the question whether psychoses might not have their etiology in the endogenous production of substances similar to LSD. Other investigators believe that LSD intoxication is an especially suitable psychosis model of schizophrenia (R. Fisher, F. Georgi, and R. Weber, Schweizerische Medizinische Wochenschri-ft, volume 81, page 817, 1951).

D. W. Woolley has demonstrated that LSD also produces abnormal behavior in mice (Proceedings of the National Academy of Science, volume 41, page 338, 1955). This was also demonstrated for rats by C. A. Winter and L. Flataker, who designed a bioassay for quantitation of antagonistic effects of drugs on the psychic action produced by LSD in rats (Proceedings of the Society for Experimental Biology and Medicine, volume 92, page 285, 1956). Using this bioassay, I. R. Bergen, D. Krus and G. Pincus demonstrated that certain steroid hormones and metabolites had biological activity in suppressing LSD-induced behavior changes in rats (Proceedings of the Society for Experimental Biology and Medicine, volume 105, page 254, 1960).

It has been found that the 2-androsten-16,l7-diol and 2-androsten-17-ol-16-one are effective in the prevention ind suppression of schizophrenic behavior induced by It is an object of this invention to provide new and useful steroids which antagonize or suppress the induction of psychotic conditions by LSD in mammals. It is a further objects to provide methods for producing such compounds by economical and efficient methods from readily available starting materials. It is a further object to produce novel 2-androsten-16,l7-diol and Z-androsten- 17-ol-16-one compounds which have useful physiological properties in antagonizing or suppressing the induction of psychotic conditions in mammals by LSD. These and other objects are apparent from and are achieved in accordance with the following disclosure.

The compounds of this invention are produced from 2-androsten-17-one [J. Biol. Chem., 136, 483 (1944)] by nitrosation with an alkyl nitrite in the presence of an alkali metal alkoxide to form 16-oximino-2-androsten-17- one followed by reaction of the oximino compound with zinc in aqueous acid to produce the corresponding l6-ketol7fl-hydroxy compound. The latter, if desired, can be reduced with an alkali metal hydride of aluminum or boron to produce a 16,8-hydroxy substituent.

The nitrosation is carried out with an excess of alkyl nitrite such as isoamyl nitrite in an excess of potassium t-butoxide in t-butyl alcohol at a temperature from 10 to 50 C., preferably at room temperature. After the reaction has been allowed to go to completion (1-4 hours) the solution is diluted with water and treated with glycine to destroy the excess of alkyl nitrite. The precipitate of the resulting l6-oximino-2-androsten-17-one is removed, purified and dried. This product can be treated in aqueous acetic acid (or similar aqueous alkanoic acids) with zinc dust to convert the 16-oximino substituent to a 16-keto substituent. This reaction is carried out at a temperature in the range of'80150 C. over a period of time from 15 hours. On dilution of the reaction mixture with water, the steroid is precipitated and can be separated by conventional procedures. The resulting 2-androsten-17B- ol-16-o'ne can then be reduced with-an alkali metal hydride of boron or aluminum such as sodium borohydride, lithium aluminum hydride or potassium borohydride in a lower alk'anol solvent at low temperature (025 C.) over a period of several hours. On dilution of the alkanol solvent with water, the desired 2-androsten-16fl,17 3-diol compound precipitates and is removed and purified by conventional procedures.

2-androsten-16oc,17B-diol is produc'edfrom 2-androsten 16-0ne-17fi-Ol by reduction of the 16-keto group with an alkali metal amalgam in the presence of a dilute acid. The reduction is conveniently carried out in alcohol solution containing sufiicient acetic acid to neutralize the alkali metal hydroxide that is formed. The reaction is preferably carried out at'low temperature (below C.) with an excess of alkali metal amalgam. The resulting 2- androsten-l 6a,l7fl-diol is isolated by extraction with an organic solvent.

The invention is disclosed in further detail by means of the following examples which are provided for purposes of illustration only. It will be understood by those skilled in the art that various modifications in operating conditions and equivalent materials can be made without departing from this invention.

EXAMPLE 1 16-oximin0-2-androstem1 7 -one To a solution of 6.5 g. of 2-androsten-17-one in 240 ml. of t-butyl alcohol containing 0.625 millimole of potassium t-butoxide per ml. was added 3.0 ml. of isoamyl nitrite. The resulting mixture was stirred for 2.5 hours. Another 3.0 ml. of isoamyl nitrite was added and stirring continued for 2.5 hours longer. The reaction mixture was poured into 3 liters of ice water containing 75 g. of glycine. Then 50 ml. of ice water containing 6 ml. of concentrated sulfuric acid was added and the mixture allowed to stand for one hour. The precipitate of 16- oximino-Z-androsten-l7-one was removed by filtration and dried in vacuum; yield 7.19 g.; M.P. 177-178 C. (decomp.). After recrystallization from 90% methanol the 16-oximino-2-androstenl7-one melted at 179-180" C. (decomp.).

EXAMPLE 2 Z-androsten-I 7 fl-ol-I 6 -one OH CH l A solution of 4.19 g. of 16-oximino-2-androsten-17-one in 230 ml. of acetic acid and 115 ml. of water was heated with steam and treated with 11.0 g. of Zinc dust and an additional 115 ml. of water. The mixture was heated to reflux for 1.75 hours. Then the solution was decanted from the zinc through a glass wool filter; the flask and zinc were washed with 20 ml. of 50% aqueous acetic acid and the wash liquor filtered through the glass wool filter and added to the prior filtrate. The combined filtrate was added slowly to 960 ml. of water and the mixture kept at 0 C. for hours! The precipitate of 2-androsten- 17fi-ol-16-one which formed was separated by filtration and dried; yield 3.08 g.; M.P. 100-102 C. After recrystallization from 75% methanol 2-androsten-l7B-ol- 16-one melted at l31.5132 C.

EXAMPLE 3 with 1.5 g. of sodium borohydride. The reaction mixture was kept at 0 C. for 2 hours and at 22 C. for one hour. The reaction mixture was poured into 1200 ml. of iced 5% sodium chloride solution and the resulting mixture was refrigerated. The precipitate (of 2-androsten- 165,17B-diol) which formed was collected on a filter and dried. It was dissolved in 20 ml. of hot methanol containing 5 drops of concentrated hydrochloric acid and 5 ml. of water, treated with activated charcoal, filtered and chilled to 25 C. The precipitate was dissolved in the minimum amount of hot 70% methanol containing 0.2 g. of potassium hydroxide, refluxed for an hour, and chilled to 25 C. After further recrystallization from absolute ethanol there was obtained 2-androsten-16fl,17fl-diol of M.P. 156 C. e EXAMPLE 4 OH CH A solution of 1.22 g. of 2-androsten-16-one-17fl-ol in 150 ml. of ethanol and 15 ml. of acetic acid was maintained at 40 C. while 300 g. of 2% sodium amalgam was added. The mixture was kept at 40 C. and gently swirled. When sodium acetate began to precipitate 10 ml. of 50% acetic acid was added, followed by 600 ml. of water. The mixture was extracted with 800 ml. of butanol and the butanol extract was washed with water, with 1 N hydrochloric acid, with water, with 1 N sodium hydroxide solution and finally with dilute sodium chloride solution. The butanol solution was evaporated to dryness and the residue of 2-androsten-16u, l7fi-diol was dissolved in acetone and treated with activated charcoal, the solution was filtered, mixed with petroleum ether, and evaporated until crystallization of 2-androsten-16m,l7fldiol began. The solution was cooled and filtered and the product was rinsed with petroleum ether and dried. 2- androsten-16u,17fl-dio1 melts at 194195 C.

I claim:

1. A steroid of the formula CHa a... n l f1 wherein X is a member of the group consisting of the carbonyl radical and the hydroxymethylene radical.

2. Z-androsten-17fl-ol-16-one.

3. Z-and-r-osten-165,17fl-dio1.

4. 2-androsten-16a,17,B-diol.

5. 16-oximino-2-androsten-17-one.

References Cited by the Examiner UNITED STATES PATENTS 3,144,447 8/ 1964 Bowers et al. 260397.4

LEWIS GOTTS, Primary Examiner. 

5. 16-OXIMINO-2-ANDROSTEN-17-ONE. 